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Pierce™ Chicken IgY Purification Kit Thermo Scientific™

The Thermo Scientific Pierce Chicken IgY Purification Kit purifies chicken antibodies from hen egg yolks using a unique delipidation reagent to separate protein from lipid and a precipitation reagent to selectively isolate the IgY fraction.

Features of the Chicken IgY Purification Kit:

More for your money—purifies twice the amount of IgY than the leading competitor's kit with a lower cost-per-mg of IgY purified
High recovery and purity—obtain 80 to 120 mg IgY per egg yolk at 85% to 95% purity by SDS-PAGE analysis
Ease-of-use—simple precipitation method purifies chicken IgY without affinity columns
Trouble-free method—fewer steps than other IgY purification kits
Flexibility—eggs can be stored in buffer and purified at a later date
Convenience—use eggs directly out of refrigerator; no need to wait for them to warm up

Typical IgG affinity resins (Protein A or Protein G) do not bind chicken IgY and therefore are not effective for IgY purification. The Pierce Chicken IgY Purification Kit uses a delipidation-and-precipitation procedure that is optimized to recover approximately 100 mg of 90% pure IgY from fresh egg yolks of appropriately immunized chickens. The resulting polyclonal IgY is sufficiently pure for direct use in many immunodetection procedures, or it can be easily affinity-purified against specific antigen because it is dissolved in PBS and thoroughly clarified.

Affinity chromatography with Protein A or Protein G cannot be used to purify IgY from chicken serum or egg yolk because this species of immunoglobulins are not recognized by the Protein A, G or L binding sites. However, the Pierce Chicken IgY Purification Kit provides a simple and reliable method for recovering large amounts of chicken IgY from egg yolk with our unique delipidation reagent that separates the proteins from lipid. (The egg yolk can be stored in the delipidation reagent a year or longer before processing.) After lipid separation, simply mix the sample with the IgY Precipitation Reagent which causes the IgY to separate from the lipid fraction during centrifugation. The precipitated chicken IgY is then easily recovered by dissolution in phosphate-buffered saline (PBS).

Chicken IgY vs. Mammalian IgG:
The terms IgG and IgY are commonly interchanged when referring to chicken immunoglobulin. Immunoglobulins from chickens and other avian species bear some resemblance to mammalian IgG, but also display some unique structural and functional characteristics that distinguish them from IgG. IgY is the accepted/proper term for chicken antibodies. Chicken IgY is the functional equivalent to mammalian IgG. It is found in the serum of chickens and is passed from the mother chicken to the embryo via the egg yolk, imparting a high concentration of chicken IgY to developing embryo.

Advantages of Chicken IgY:
Chicken IgY immunoglobulins can be harvested from the egg yolk as well as serum and blood. There are many advantages to using these antibodies:

• Unlike mice, rabbits or other mammals, hens elicit a strong antibody response against highly conserved mammalian protein sequences
• Chicken antibodies can be obtained without sacrificing or bleeding the animal—simply collect the eggs and use the Pierce Chicken IgY Purification Kit
• Each yolk can yield 100-150 mg of antibody.
• Chicken IgY has less cross-reactivity toward mammalian proteins—does not bind Fc receptors!

Imject™ Freund's Incomplete Adjuvant (FIA) Thermo Scientific™

Thermo Scientific Imject Freund's Incomplete Adjuvant is a popular nonspecific stimulator of the immune response for use in antibody production immunization protocols.

This product comes in 10 mL ampules that are convenient and ready-to-use. The water-and-oil emulsion greatly enhances immune responses to immunogens when mixed and injected with the prepared antigen, and is typically used for booster injections. The ampules comprise conveniently sized aliquots that provide long shelf life.

Highlights of Freund's Incomplete Adjuvant (FIA):
• Also called Incomplete Freund's Adjuvant (IFA)
• Water-in-oil emulsion without Mycobacterium
• Enhances immune response in booster injections
• Typically used for subsequent boosts after initial injection with FCA
• Disadvantage: difficult to mix with immunogen

Adjuvants are nonspecific stimulators of the immune response. When mixed with an antigen or immunogen, adjuvants help to deposit or sequester the injected material thereby helping to increase antibody response. Adjuvants enhance the immune response to compounds that are already immunogenic; they do not confer immunogenicity to non-immunogenic haptens. To make prospective antigens more immunogenic, it is necessary to conjugate them to a carrier protein or some other complex, immunogenic molecule.

For added safety when opening glass ampules, consider using our Ampule Breakers, which are disposable safety devices that protect the fingers when breaking open a glass ampule.

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Imject™ Alum Adjuvant
Imject™ Freund's Complete Adjuvant (FCA)

Easy-Titer™ Rabbit IgG Assay Kit Thermo Scientific™

The Thermo Scientific Easy-Titer Rabbit IgG Assay Kit includes antibody-sensitized microspheres to measure the specific concentration of antibodies by an easy and rapid microagglutination technique using standard microplates and UV-Vis plate reader (spectrophotometer). This kit is specific for rabbit IgG and, unlike total protein assays, can specifically measure the concentration of target antibody in samples (e.g., serum, plasma, culture supernatant) that contain other proteins. It is sensitive, requiring very small sample volumes. Antibody concentration is determined from the assay response (absorbance) by comparison to a standard curve prepared using dilutions of a known antibody sample (sold separately).

General features of Easy-Titer Antibody Assay Kits:

Antibody-based specificity—measure concentration of target antibody in a sample, not just total protein; no need to purify antibody to assess its concentration
Faster and easier than ELISA—three-component, homogenous assay; 10 minutes total incubation time
No special equipment needed—uses standard vortex mixer, pipetter, 96-well microplate, plate shaker and reader (measure absorbance at 340nm or 400nm)
Sensitive—assay range (standard curve) 8 to 500 ng/mL; use sample at 15 to 300 ng/mL for optimal results
Reproducible—coefficient of variation < 5%; error depends on dilution and pipetting technique
Antibody standards sold separately—see product list for suggested products; use any antibody standard with proper target identity and known concentration (greater than 10 µg/mL)
Kits for five popular targets—choose a kit specific for a particular species and class of immunoglobulin; no cross-reaction with other species and classes of the target antibody

Easy-Titer Assay Kits detect and measure specific target antibodies using agglutination of microspheres that are coated ('sensitized') with the specific anti-IgG or IgM polyclonal antibodies. In the appropriate aqueous buffer (supplied in kit), the monodispersed antibody-coated microspheres (> 1 µM diameter) have highest absorptivity (λ-max) to incident light having a wavelength (340nm) that is equal to approximately half their diameter. When sample is added, two or more microspheres bind to each antibody target via their coated specific polyclonal antibodies, and this agglutination into effectively larger apparent spheres results in proportional decrease in absorptivity (lower absorbance).

Typical microagglutination assays depend on a change in light-scattering and corresponding change in transmittance, to which absorbance is inversely related. Easy-Titer Assay Kits use a special dilution buffer whose refractive index eliminates the effect of light-scattering on the monodispersed microspheres for the measurement wavelength used. Because of this, the final 10- to 20-fold dilution of the sample for use in the assay must be done using the Dilution Buffer supplied in the kit.

Easy-Titer Antibody Assays are faster and easier than ELISA:
• Prepare standards (5 to 500 ng/mL) by diluting purified antibody in Kit Dilution Buffer.
• Prepare samples by diluting in Dilution Buffer to within assay range (8 to 500 ng/mL).
• Vortex vial of microsphere beads to create homogeneous suspension.
• Pipette 20 µL of bead suspension and 20 µL of each sample and standard into 96-well microplate wells.
• Incubate microplate for 5 minutes with vigorous mixing.
• Add 100 µL of Kit Blocking Reagent.
• Incubate microplate for 5 minutes with vigorous mixing.
• Measure absorbance on standard plate reader (340nm or 405nm).
• Plot standard curve and interpolate samples to determine concentration.

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Easy-Titer™ Mouse IgG Assay Kit
Easy-Titer™ Human IgG (H+L) Assay Kit
Easy-Titer™ Human IgG (gamma chain) Assay Kit
Easy-Titer™ Human IgM Assay Kit

MBP Column Preparation Buffer for Pierce™ IgM Purification Kit Thermo Scientific™

MBP Column Preparation Buffer, formulation: Tris buffer with sodium chloride, sodium azide and EDTA; pH 7.4. For use with Pierce Mannan Binding Protein Agarose, and the Pierce IgM Purification Kit.

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Elution Buffer for Pierce™ IgM Purification Kit

Easy-Titer™ Human IgG (gamma chain) Assay Kit Thermo Scientific™

The Thermo Scientific Easy-Titer Human IgG (gamma chain) Assay Kit includes antibody-sensitized microspheres to measure the specific concentration of antibodies by an easy and rapid microagglutination technique using standard microplates and UV-Vis plate reader (spectrophotometer). This kit is specific for the human IgG gamma chain and, unlike total protein assays, can specifically measure the concentration of target antibody in samples (e.g., serum, plasma, culture supernatant) that contain other proteins. It is sensitive, requiring very small sample volumes. Antibody concentration is determined from the assay response (absorbance) by comparison to a standard curve prepared using dilutions of a known antibody sample (sold separately).

General features of Easy-Titer Antibody Assay Kits:

Antibody-based specificity—measure concentration of target antibody in a sample, not just total protein; no need to purify antibody to assess its concentration
Faster and easier than ELISA—three-component, homogenous assay; 10 minutes total incubation time
No special equipment needed—uses standard vortex mixer, pipetter, 96-well microplate, plate shaker and reader (measure absorbance at 340nm or 400nm)
Sensitive—assay range (standard curve) 8 to 500 ng/mL; use sample at 15 to 300 ng/mL for optimal results
Reproducible—coefficient of variation < 5%; error depends on dilution and pipetting technique
Antibody standards sold separately—see product list for suggested products; use any antibody standard with proper target identity and known concentration (greater than 10 µg/mL)
Kits for five popular targets—choose a kit specific for a particular species and class of immunoglobulin; no cross-reaction with other species and classes of the target antibody

Easy-Titer Assay Kits detect and measure specific target antibodies using agglutination of microspheres that are coated ('sensitized') with the specific anti-IgG or IgM polyclonal antibodies. In the appropriate aqueous buffer (supplied in kit), the monodispersed antibody-coated microspheres (> 1 µM diameter) have highest absorptivity (λ-max) to incident light having a wavelength (340nm) that is equal to approximately half their diameter. When sample is added, two or more microspheres bind to each antibody target via their coated specific polyclonal antibodies, and this agglutination into effectively larger apparent spheres results in proportional decrease in absorptivity (lower absorbance).

Typical microagglutination assays depend on a change in light-scattering and corresponding change in transmittance, to which absorbance is inversely related. Easy-Titer Assay Kits use a special dilution buffer whose refractive index eliminates the effect of light-scattering on the monodispersed microspheres for the measurement wavelength used. Because of this, the final 10- to 20-fold dilution of the sample for use in the assay must be done using the Dilution Buffer supplied in the kit.

Easy-Titer Antibody Assays are faster and easier than ELISA:
• Prepare standards (5 to 500 ng/mL) by diluting purified antibody in Kit Dilution Buffer.
• Prepare samples by diluting in Dilution Buffer to within assay range (8 to 500 ng/mL).
• Vortex vial of microsphere beads to create homogeneous suspension.
• Pipette 20 µL of bead suspension and 20 µL of each sample and standard into 96-well microplate wells.
• Incubate microplate for 5 minutes with vigorous mixing.
• Add 100 µL of Kit Blocking Reagent.
• Incubate microplate for 5 minutes with vigorous mixing.
• Measure absorbance on standard plate reader (340nm or 405nm).
• Plot standard curve and interpolate samples to determine concentration.

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Easy-Titer™ Mouse IgG Assay Kit
Easy-Titer™ Rabbit IgG Assay Kit
Easy-Titer™ Human IgG (H+L) Assay Kit
Easy-Titer™ Human IgM Assay Kit

Imject™ Maleimide-Activated Blue Carrier™ Protein Thermo Scientific™

Thermo Scientific Imject Maleimide-Activated Blue Carrier Protein enables conjugation of sulfhydryl-containing peptide antigens to this soluble immunogenic carrier protein for immunization and effective antibody production.

Features of Imject Maleimide-Activated Blue Carrier Protein:

Optimized activation—specifications require that each lot is activated with more than 400 moles of sulhydryl-reactive maleimide per mole of carrier, ensuring consistently high levels of conjugation
Sulfhydryl-reactive—engineered for conjugating cysteine-containing peptide antigens to make effective immunogens for high-titer antibody production
Simple, rapid procedure—complete the hapten-carrier conjugation reaction in two hours to yield sufficient immunogen for several injections and booster immunizations of host animals
High-quality carrier protein—Blue Carrier Protein is ultrapure Concholepas concholepas hemocyanin (CCH) prepared from molluscan hemolymph
Highly immunogenic and soluble—CCH has a high molecular mass that elicits a stronger immune response than BSA or ovalbumin

Blue Carrier* Protein is a large hemocyanin that is highly immunogenic like keyhole limpet hemocyanin (KLH) yet is more soluble. The carrier is rendered sulfhydryl-reactive by chemical modification with Sulfo-SMCC crosslinker, and the resulting maleimide-activated protein is then stabilized and lyophilized in phosphate buffer. Maleimide groups form covalent crosslinks with sulfhydryl (-SH) moieties on cysteine residues of peptides and other thiol-containing haptens. Numerous antigen molecules can be conjugated to each Blue Carrier Protein molecule to make them highly immunogenic and increase the likelihood of eliciting immunized animals to produce high-titer antisera to the epitopes of interest.

Carrier proteins are large, complex molecules capable of stimulating an immune response upon injection. Successful production of antibodies specific to small antigens (i.e., peptides or drug compounds) requires that these haptens be covalently conjugated to a larger, more complex molecule (usually a protein) to make them immunogenic. Carrier proteins are chosen based on immunogenicity, solubility, and whether an adequate level of hapten-carrier conjugation can be achieved.

Blue Carrier* Protein is specially purified hemocyanin from the mollusk Concholepas concholepas. The CCH protein is composed of two very large polypeptide subunits (404/351 kDa) that form an extremely stable heterodidecameric structure even in the absence of divalent cations. (By contrast, KLH has a less stable and soluble homodidecameric structure). The complex molecular arrangement of CCH subunits contains diverse repeated antigenic structures that elicit a strong immune reaction mediated by T and B lymphocytes.

Because of their large size and molecular complexity, KLH and CCH hemocyanins are carrier proteins of choice for use as immunogens to produce antibodies against haptens and peptides. Moreover, studies suggest that the strong DTH immune response elicited by hemocyanins in animals and in humans has beneficial therapeutic effects in certain types of cancer. New developments in the immunotherapy of cancer have taken advantage of the unique immunogenic properties of hemocyanins in the development of novel conjugate vaccines for treatment of emerging diseases.

More Product Data
Carrier protein activation and conjugation data for immunogen preparation

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Imject™ Maleimide-Activated Blue Carrier™ Protein

Imject™ cBSA (in PBS) Thermo Scientific™

Thermo Scientific Imject Cationized Bovine Serum Albumin (cBSA) functions as an easy-to-use and highly immunogenic carrier protein for preparing antigens for use in antibody production.

Imject cBSA is prepared by modifying native BSA with excess ethylenediamine, essentially capping all negatively-charged carboxyl groups with positively-charged primary amines. The result is a highly positively-charged protein (pI > 11) that has significantly increased immunogenicity compared to native BSA. In addition, the increased number of primary amines provides for a greater number of antigen molecules to be conjugated with typical crosslinking methods. Imject cBSA is an excellent choice for immunogen preparation when greater immunogenicity is needed relative to BSA but conditions are not amenable to using keyhole limpet hemocyanin (KLH) or Blue Carrier Protein.

Features of the Imject Cationized Bovine Serum Albumin:

• Purified bovine serum albumin (BSA) that is cationized and supplied lyophilized in PBS
• Single polypeptide protein with molecular mass > 67,000 Daltons
• Contains 59 lysine residues, plus numerous other primary amines added by cationization, most of which are capable of reacting with a conjugation reagent to attach antigens
• Net positive charge (pI >11), conferring solubility in conditions not suitable for the negatively-charged native BSA (pI 5.1)
• Significantly greater immunogenicity than native BSA because the positively charged protein is processed differently by the immune-response system (see references).

Carrier proteins are large, complex molecules capable of stimulating an immune response upon injection. Successful production of antibodies specific to small antigens (i.e., peptides or drug compounds) requires that these haptens be covalently conjugated to a larger, more complex molecule (usually a protein) to make them immunogenic. Carrier proteins are chosen based on immunogenicity, solubility, and whether adequate conjugation with the carrier can be achieved. Imject Cationized BSA (cBSA), which was previously called "SuperCarrier Protein" is very soluble in aqueous buffers and contains numerous primary amines that can be targeted for conjugation with glutaraldehyde, EDC, and other crosslinking reagents. For sulfhydryl-target conjugation (e.g., cysteine-containing peptide antigens), use Imject Maleimide-Activated cBSA.

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Pierce™ Rapid Antibody Isotyping Kit - Mouse Thermo Scientific™

The Thermo Scientific Pierce Rapid Mouse Antibody Isotyping Kit is a single-use, cassette-based kit for 5-minute lateral-flow assay determination of mouse monoclonal antibody class and subclass identity.

This antibody isotyping kit uses cartridges that provide a color-readout of the monoclonal antibody isotype within five minutes after pipetting a small amount of diluted culture supernatant or ascites fluid sample into the sample well. The kit determines mouse IgG1, IgG2a, IgG2b, IgG3, IgA, IgM.

Features of the Pierce Rapid Mouse Antibody Isotyping Kit:

Long shelf life—stable for at least one year at room temperature
Single-step—add diluted antibody sample to the loading-well of cassette
Fast—within 5 minutes a band appears that indicates the antibody isotype
Sensitive—detect antibodies in tissue culture media or ascites fluid samples at any concentration greater than 10 ng/mL
Specific—assay does not cross-react with fetal bovine serum (FBS)
Reliable—results are similar to the standard ELISA-based isotyping assays

Each Pierce Rapid Isotyping Kit assay is performed by simply adding a properly diluted tissue culture supernatant or mouse ascites sample to the well of the small cassette. Gold conjugates embedded in the cassette form specific class- and subclass-soluble complexes with the antibodies in the sample. These complexes travel the length of the membrane and are resolved on the anti-isotype and class-specific antibody-impregnated membrane. Results are displayed as a red band indicating the antibody isotype or subclass.

Determining the class and subclass of a monoclonal antibody is useful in planning the best immunoglobulin purification method. For example, mouse IgA and IgM are best purified by size (i.e., gel exclusion) or using immunoaffinity separation columns. Mouse IgG2a and IgG2b are purified with immobilized Protein A at pH 7-8, while Mouse IgG1 binds best to Protein A at pH 8-9. Immunoglobulin that contains kappa light chains can be purified using immobilized Protein L.

The Pierce Rapid Isotyping Kit is compatible with both tissue culture supernatant and mouse ascites fluid, and is more sensitive than conventional latex-bead based dipstick assays and much faster than ELISA-based isotyping assays. The kit requires only 0.5µL ascites fluid, 5 to 55µL of cell culture supernatant or 1.5ng of purified antibody.

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Imject™ Maleimide PEGylated mcKLH Thermo Scientific™

Thermo Scientific Imject Maleimide PEGylated mcKLH enables conjugation of sulfhydryl-containing peptide haptens to the highly immunogenic keyhole limpet hemocyanin (KLH) carrier protein for immunization and antibody production. This PEGylated form offers even greater hydrophilicity and solubility than typical forms.

Features of Imject Maleimide-Activated mcKLH:

Optimized activation—specifications require that each lot is activated with more than 400 moles of sulhydryl-reactive maleimide per mole of KLH, ensuring consistently high levels of conjugation
Sulfhydryl-reactive—engineered for conjugating cysteine-containing peptide antigens to make effective immunogens for high-titer antibody production
Simple, rapid procedure—complete the hapten-carrier conjugation reaction in two hours to yield sufficient immunogen for several injections and booster immunizations of host animals
High-quality carrier protein—KLH is isolated from the mollusk Megathura crenulata (keyhole limpet); mcKLH is a high-quality form of KLH that is harvested from limpets grown in mariculture
Highly immunogenic—KLH has a high molecular mass (4.5 x 105 to 1.3 x 107 Daltons; aggregates of 350 and 390 kDa subunits) and elicits a stronger immune response than BSA or ovalbumin
Excellent solubility—our preparations of mariculture KLH are more soluble than typical forms

The high-quality mariculture KLH is rendered sulfhydryl-reactive by chemical modification with Sulfo-SMCC crosslinker, and the resulting maleimide-activated protein is then stabilized and lyophilized in phosphate buffer. Maleimide groups form covalent crosslinks with sulfhydryl (-SH) moieties on cysteine residues of peptides and other thiol-containing haptens. Numerous antigen molecules can be conjugated to each KLH molecule to make them highly immunogenic and increase the likelihood of eliciting immunized animals to produce high-titer antisera to the epitopes of interest. Select from among several package sizes of activated KLH protein, or choose the complete conjugation kit that includes optimized buffers and trouble-free spin desalting columns.

Carrier proteins are large, complex molecules capable of stimulating an immune response upon injection. Successful production of antibodies specific to small antigens (i.e., peptides or drug compounds) requires that these haptens be covalently conjugated to a larger, more complex molecule (usually a protein) to make them immunogenic. Carrier proteins are chosen based on immunogenicity, solubility, and whether an adequate level of hapten-carrier conjugation can be achieved.

Keyhole limpet hemocyanin (KLH) is widely used as a carrier protein for conjugation to haptens and other antigens to make them more immunogenic for the purpose of antibody production. Because of its large mass and complexity, KLH elicits a stronger immune response than other carrier proteins. Because the protein is derived from a mollusk, it is phylogenetically distant from mammalian species and less likely to produce antibodies that will cross-react with typical target samples in assays.

More Product Data
Carrier protein activation and conjugation data for immunogen preparation

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Imject™ Maleimide-Activated mcKLH
Imject™ Maleimide-Activated mcKLH Spin Kit
Imject™ Maleimide Conjugation Buffer

Melon™ Gel Chromatography Cartridges, 1 mL Thermo Scientific™

The Thermo Scientific Melon Gel Chromatography Cartridge is a convenient, ready-to-use prepacked chromatography column for the isolation and purification of antibodies. Our Melon Gel IgG Purification Resin offers a simpler and faster alternative to traditional Protein A and Protein G methods for purification of human, mouse and rabbit IgG from serum and other sources.

Features of Melon Gel IgG Purification Resin:

Simple, one-step protocol—IgG purification with Melon Gel Resin does not involve binding, washing, and multiple elution steps required by Protein A and Protein G
Rapid purification—antibody purification from serum is four to six times faster than Protein A or G methods
High recovery and purity—for many species of serum antibody recovery is greater than 90% with purity greater than 80%
Minimal dilution—reduced need to concentrate antibody before storage or use
Gentle purification—No harsh antibody elution conditions helps retain IgG activity
Regenerable support—Melon Gel Resin can be used for multiple antibody purifications

These cartridges are compatible with major automated liquid-chromatography systems or manual syringe processing, and attach directly to ÄKTA™ or FPLC Systems without additional connectors. An accessory pack, included with each product, readily adapts cartridges for use with Luer-Lok Syringe Fittings or 1/16” tubing. The Melon Gel Cartridges provide fast, easy and reproducible chromatographic separations and can be regenerated for multiple uses.

Melon Gel IgG Purification Resin enables purification of antibodies from serum and other sample types. Melon Gel Resin binds and removes serum proteins, thereby allowing pure IgG to be collected in the flow-through fraction. The procedure is gentle because no harsh elution conditions are involved.

Melon Gel Resin contains a proprietary ligand that retains most protein found in serum, ascites and culture supernatants, while allowing IgG to pass through the support and be collected in the flow-through fraction. Because Melon Gel Resin is not a bind-and-release support, it is extremely fast and gentle to ant ibodies, resulting in antibody preparations of high purity and high activity.

Melon Gel Resin works with antibodies from a variety of species and subclasses, many of which do not purify efficiently with Protein A or Protein G. The resulting recovery and purity of the IgG isolated by this method rivals that obtained from the same samples using bind-and-release supports such as Protein A or Protein G. Furthermore, antibodies purified with Melon Gel Resin are recovered ready for use in immunoassays or subsequent conjugation or modification steps; there is no need to neutralize or desalt the recovered antibody to remove harsh elution buffers.

Melon Gel IgG Purification saves time compared to classical Protein A or Protein G methods without compromising the quality of the antibody. Compared to antibody affinity purification with Protein A or Protein G, Melon Gel IgG Purification is simpler, faster and provides greater yields and higher purity for a wider variety of antibodies species.

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CaptureSelect™ IgA Affinity Matrix Thermo Scientific™

The CaptureSelect™ IgA Affinity Matrix has specifically been designed for the purification of human IgA from recombinant and plasma sources.

Features of this affinity matrix include:

• Platform for all human IgA, binding to IgA1, IgA2 , dimeric IgA, and S-IgA (secretory IgA)
• Excellent scalability
• Non-animal-derived

Free of Animal Components
Our CaptureSelect™ products are affinity ligands created by a proprietary technology based on Camelid-derived single domain antibody fragments. The ligand is a 13 kDa single domain fragment comprising the 3 CDRs that form the antigen binding domain, efficiently produced by the yeast Saccharomyces cerevisiae in a production process free of animal components.

Main Characteristics:

Matrix: agarose-based, aldehyde-activated
Average particle size: 65 ± 10 µm
Ligand: CaptureSelect™ IgA affinity ligand
Ligand coupling method: aldehyde coupling via NH2 residues of the ligand
Binding capacity: ~8 g/L resin
Elution conditions: 0.1 M glycine, pH 3.0
Flow characteristics: 200–400 cm/h
Formulation buffer: 20%(v/v) ethanol

Imject™ Maleimide-Activated Ovalbumin Thermo Scientific™

Thermo Scientific Imject Maleimide-Activated Ovalbumin is sulfhydryl-reactive ovalbumin (OVA) for hapten-carrier conjugation with cysteine-containing peptides and other thiolated antigens.

Features of Imject Maleimide-Activated Ovalbumin:

• Maleimide-Activated Ovalbumin, supplied lyophilized in PBS (pH 7.2) with stabilizer
• Activated with 5 to 15 moles of maleimide per mole of ovalbumin
• Ideal for conjugating cysteine-containing peptide antigens to make immunogen or screening target
• Conjugation reaction complete in 2 hours; results in stable, covalent thioether bonds
• Single polypeptide protein with molecular mass of 45,000 Daltons
• Contains 14 aspartic acid and 33 glutamic acid residues whose carboxyl groups account for the net negative charge (pI 4.63) of OVA
• Not as immunogenic as keyhole limpet hemocyanin (KLH) but more soluble in DMSO (dimethyl sulfoxide) for applications that require this solvent
• Useful as an irrelevant protein carrier for antibody screening and immunoassays after using KLH as the carrier protein to generate the immune response against the hapten

This product consists of purified, carrier-grade ovalbumin that has been modified with SMCC crosslinker to attach maleimide groups capable of forming covalent crosslinks with sulfhydryl (-SH) moieties on cysteine residues of peptides and other molecules. Immunogens are easily prepared from peptide antigens that have been engineered with a terminal cysteine residue as a precise conjugation point. Several antigen molecules can be attached per carrier protein molecule without blocking intended epitopes by crosslinking to primary amines or carboxylates within the peptide sequence.

Carrier proteins are large, complex molecules capable of stimulating an immune response upon injection. Successful production of antibodies specific to small antigens (i.e., peptides or drug compounds) requires that these haptens be covalently conjugated to a larger, more complex molecule (usually a protein) to make them immunogenic. Carrier proteins are chosen based on immunogenicity, solubility, and whether adequate conjugation with the carrier can be achieved.

Ovalbumin (OVA), the most abundant protein in chicken egg whites, is a convenient protein for a variety of uses in the laboratory. The 45kDa protein is sufficiently large and complex to be mildly immunogenic. Consequently, OVA can be used as a carrier protein for conjugation to haptens and other antigens to make them more immunogenic for immunization. If not commonly used to prepare the primary immunogen, OVA is a popular carrier protein to make antigens more amenable to analysis (e.g., screening assays involving plate coating) of antibodies that were produced using KLH or other more immunogenic carrier protein.

Imject Ovalbumin is not as immunogenic as other proteins, such as keyhole limpet hemocyanin (KLH). However, OVA's independent structure make it especially useful as a carrier for peptides and other small antigens that by themselves are difficult to manipulate in typical screening assays such as ELISA. Although it is more hydrophobic and less soluble in aqueous buffers than BSA, ovalbumin remains soluble in up to 70% DMSO, making it useful for conjugation with haptens that require DMSO for solubility. Ovalbumin contains numerous primary amines, some of which have been modified to make Maleimide-Activated Ovalbumin.

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Pierce™ Fab Preparation Kit Thermo Scientific™

Thermo Scientific Pierce Fab Preparation Kit uses immobilized papain protease to digest human or mouse IgG antibodies to make separate Fab and Fc fragments and subsequently to purify the Fab using Protein A agarose.

Features of the Fab Preparation Kit:

High capacity—use the standard size kit for fragmenting 0.25 to 4 mg IgG.
Enzyme-free digestion products—Immobilized Papain (beaded agarose resin) provides for control of the digestion reaction and complete removal of resulting antibody fragments from the proteolytic enzyme
Suitable for human and other species of IgG—the kit procedure is optimized for human, mouse and rabbit IgG. Papain-based digestion is effective for certain other species and subclasses of IgG (e.g., goat and pig), although the purification step requires that the antibody effectively binds to Protein A. (Note: for best results with mouse IgG1, use Part No. 44980.)
Provides ready-to-use Fab—digestion and final recovery of purified Fab fragments occurs in neutral pH sodium phosphate buffer, suitable for storage or immediate use in typical applications
Complete—kits include all reagents needed to prepare and purify antibody fragments
Fast—spin format greatly reduces sample processing time
Flexible—Protocols are included for multiple species and IgG subclasses, as well as sample size and concentration.
Efficient—enhanced yield and sample purity

These Fab Preparation Kits are suitable for human, rabbit, mouse and other species and subclasses of IgG. The papain antibody digestion reaction is performed in convenient disposable spin columns that allow efficient removal of the immobilized protease and maximum recovery of the IgG fragments. Also included in the kits are Thermo Scientific NAb Protein A Spin Columns and buffers to efficiently purify the resulting fragments. Protein A binds the Fc fragments and undigested IgG, allowing the pure Fab fragments to be recovered in the flow-through fraction. The kits also include Thermo Scientific Zeba Desalting Spin Columns for preparing the IgG sample quickly without dilution instead of utilizing time-consuming dialysis steps.

The kits use papain, a nonspecific thiol-endopeptidase, to enzymatically cleave whole IgG just above the hinge region to create two separate Fab fragments and one Fc fragment per antibody molecule. Because the papain protease is supplied in immobilized form as beaded agarose resin, the digestion reaction is easily stopped by removing the resin from the IgG solution; the result is digest products that are enzyme-free.

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Imject™ Purification Buffer Salts Thermo Scientific™

Thermo Scientific Imject Purification Buffer Salts, formulation: Dry powder for dissolution in 60 mL water to make 0.083M sodium phosphate, 0.9M sodium chloride, 0.1M sorbitol; pH 7.2. Sufficient for preparing 60 mL of solution for buffer-exchange of immunogens following conjugation.

More Product Data
Carrier protein activation and conjugation data for immunogen preparation

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Melon™ Gel IgG Spin Purification Kit Thermo Scientific™

The Thermo Scientific Melon Gel IgG Spin Purification Kit offers a simpler and faster alternative to traditional Protein A and Protein G methods for purification of human, mouse and rabbit IgG from serum and other sources.

Features of Melon Gel IgG Purification Kits:

Simple, one-step protocol—IgG purification with Melon Gel Resin does not involve binding, washing, and multiple elution steps required by Protein A and Protein G
Rapid purification—antibody purification from serum is four to six times faster than Protein A or G methods
High recovery and purity—for many species of serum antibody recovery is greater than 90% with purity greater than 80%
Minimal dilution—reduced need to concentrate antibody before storage or use
Robust purification—Melon Gel IgG Purification Kits work with a wide range of antibodies including many that do not purify well on Protein A or Protein G
Gentle purification—No harsh antibody elution conditions helps retain IgG activity
Regenerable support—Melon Gel Resin can be used for multiple antibody purifications

Melon Gel IgG Purification Kits enable purification of antibodies from serum and other sample types. Melon Gel Resin binds and removes serum proteins, thereby allowing pure IgG to be collected in the flow-through fraction. The procedure is gentle because no harsh elution conditions are involved.

Melon Gel Resin contains a proprietary ligand that retains most protein found in serum, ascites and culture supernatants, while allowing IgG to pass through the support and be collected in the flow-through fraction. Because Melon Gel Resin is not a bind-and-release support, it is extremely fast and gentle to antibodies, resulting in antibody preparations of high purity and high activity.

Melon Gel Resin works with antibodies from a variety of species and subclasses, many of which do not purify efficiently with Protein A or Protein G. The resulting recovery and purity of the IgG isolated by this method rivals that obtained from the same samples using bind-and-release supports such as Protein A or Protein G. Furthermore, antibodies purified with Melon Gel Resin are recovered ready for use in immunoassays or subsequent conjugation or modification steps; there is no need to neutralize or desalt the recovered antibody to remove harsh elution buffers.

Melon Gel IgG Purification saves time compared to classical Protein A or Protein G methods without compromising the quality of the antibody. Compared to antibody affinity purification with Protein A or Protein G, Melon Gel IgG Purification is simpler, faster and provides greater yields and higher purity for a wider variety of antibodies species.

Related Products
Melon™ Gel IgG Purification Kit
Melon™ Gel Purification Buffer
Melon™ Gel Regenerant
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